Digital Detection of Multiple Minority Mutants and Expression Levels of Multiple Colorectal Cancer-Related Genes Using Digital-PCR Coupled with Bead-Array
Fig 3
Specificity of multiplex ligation-dependent probe amplification-digital amplification coupled with hydrogel bead-array for detecting gene expressions of colorectal cancer (CRC)-related genes (cox-2, CD44v6, C-myc, N-ras, H-ras, and RAB5A), expression of β-actin, mutants (MUTs), and wild-type (WT) targets.
The samples were 100% targets of β-actin (A); 100% targets of CRC-related genes (B); 100% WT DNA of codon 1338 in normal tissue (C); and 100% MUT DNA of codon 1338 in SW480 cells (D). Seven pairs of probes of CRC-related genes and β-actin was added to the MLPA reaction mixture (A and B), and all probes of codons 1406, 1338 and 1356 were used to the MLPA reactions (C and D). After emulsion PCR, both the Cy5-labeled and Cy3-labeled probes were added to the bead-array for hybridization.